Functional complementation of a genetic deficiency with human artificial chromosomes.
We have shown functional complementation of a genetic deficiency in human cultured cells, using artificial chromosomes derived from cloned human genomic fragments. A 404-kb human-artificial-chromosome (HAC) vector, consisting of 220 kb of alphoid DNA from the centromere of chromosome 17, human telom...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
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2001
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| _version_ | 1797077778767544320 |
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| author | Mejía, J Willmott, A Levy, E Earnshaw, W Larin, Z |
| author_facet | Mejía, J Willmott, A Levy, E Earnshaw, W Larin, Z |
| author_sort | Mejía, J |
| collection | OXFORD |
| description | We have shown functional complementation of a genetic deficiency in human cultured cells, using artificial chromosomes derived from cloned human genomic fragments. A 404-kb human-artificial-chromosome (HAC) vector, consisting of 220 kb of alphoid DNA from the centromere of chromosome 17, human telomeres, and the hypoxanthine guanine phosphoribosyltransferase (HPRT) genomic locus, was transferred to HPRT-deficient HT1080 fibrosarcoma cells. We generated several cell lines with low-copy-number, megabase-sized HACs containing a functional centromere and one or possibly several copies of the HPRT1 gene complementing the metabolic deficiency. The HACs consisted of alternating alphoid and nonalphoid DNA segments derived only from the input DNA (within the sensitivity limits of FISH detection), and the largest continuous alphoid segment was 158-250 kb. The study of both the structure and mitotic stability of these HACs offers insights into the mechanisms of centromere formation in synthetic chromosomes and will further the development of this human-gene-transfer technology. |
| first_indexed | 2024-03-07T00:22:53Z |
| format | Journal article |
| id | oxford-uuid:7d24e734-6128-4e37-ba95-ef5ce7c11ae9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T00:22:53Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:7d24e734-6128-4e37-ba95-ef5ce7c11ae92022-03-26T21:01:45ZFunctional complementation of a genetic deficiency with human artificial chromosomes.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:7d24e734-6128-4e37-ba95-ef5ce7c11ae9EnglishSymplectic Elements at Oxford2001Mejía, JWillmott, ALevy, EEarnshaw, WLarin, ZWe have shown functional complementation of a genetic deficiency in human cultured cells, using artificial chromosomes derived from cloned human genomic fragments. A 404-kb human-artificial-chromosome (HAC) vector, consisting of 220 kb of alphoid DNA from the centromere of chromosome 17, human telomeres, and the hypoxanthine guanine phosphoribosyltransferase (HPRT) genomic locus, was transferred to HPRT-deficient HT1080 fibrosarcoma cells. We generated several cell lines with low-copy-number, megabase-sized HACs containing a functional centromere and one or possibly several copies of the HPRT1 gene complementing the metabolic deficiency. The HACs consisted of alternating alphoid and nonalphoid DNA segments derived only from the input DNA (within the sensitivity limits of FISH detection), and the largest continuous alphoid segment was 158-250 kb. The study of both the structure and mitotic stability of these HACs offers insights into the mechanisms of centromere formation in synthetic chromosomes and will further the development of this human-gene-transfer technology. |
| spellingShingle | Mejía, J Willmott, A Levy, E Earnshaw, W Larin, Z Functional complementation of a genetic deficiency with human artificial chromosomes. |
| title | Functional complementation of a genetic deficiency with human artificial chromosomes. |
| title_full | Functional complementation of a genetic deficiency with human artificial chromosomes. |
| title_fullStr | Functional complementation of a genetic deficiency with human artificial chromosomes. |
| title_full_unstemmed | Functional complementation of a genetic deficiency with human artificial chromosomes. |
| title_short | Functional complementation of a genetic deficiency with human artificial chromosomes. |
| title_sort | functional complementation of a genetic deficiency with human artificial chromosomes |
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